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. 2021 Apr;43(4):933-944.
doi: 10.1007/s10529-021-03078-1. Epub 2021 Jan 29.

MPH-GST sensing microplate for easy detection of organophosphate insecticides

Witsanu Rapichai  1 , Sudarat Chaichalerm  2 , Jhirat Mearnchu  3 , Jesdawan Wichitwechkarn  4
Affiliations

MPH-GST sensing microplate for easy detection of organophosphate insecticides

Witsanu Rapichai et al. Biotechnol Lett. 2021 Apr.

Abstract

Objective: To develop a convenient and efficient means for organophosphate (OP) insecticide detection, a simple, cost-effective, and easy-to-use absorbance-based sensing device was generated using methyl parathion hydrolase fused with glutathione-S-transferase (MPH-GST) covalently immobilized onto a chitosan film-coated microplate.

Results: With methyl parathion (MP) as a representative substrate, this MPH-GST sensing microplate had the detection limit of 0.1 µM and the linear range of 0.1-50 µM. Despite its highest stability at 4 °C, it was considerably stable at 25 °C with high activity for 30 days. It was also most stable at pH 8.0 and could be efficiently reused up to 100 rounds. The device revealed a high percentage of recovery for tap water spiked with a known concentration of MP, which was also comparable to the result obtained from gas chromatography-mass spectrometry. It also showed a high recovery of 82-100% with MP spiked agricultural products and satisfactory results with non-spiked samples. This immobilized enzyme sensing system was more sensitive and efficient than the whole cell system from our previous work.

Conclusions: All of the advantages of the MPH-GST sensing microplate developed have rendered it suitable for rapid and convenient OP screening, and for being a bio-element for fabricating a potential optical biosensor in the future.

Keywords: Covalent immobilization; MPH-GST; Methyl parathion hydrolase; Organophosphate; Sensing microplate.

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